Histochemistry

, Volume 80, Issue 6, pp 597–601 | Cite as

Basis for the specificity of anti-5-HT-like antisera in immunocytochemistry applied to the central nervous system

  • S. Peressini
  • A. Brusco
  • J. Pecci Saavedra
Article

Summary

Recently (Pecci Saavedra et al. 1982; Brusco et al. 1982, 1983) we have showed that the actual specificity of the rabbit anti-5-HT antibodies, is for the β-carboline derivatives of 5-HT as a result of cyclization of the lateral chain. We explained this as resulting from the use of formaldehyde which acted both as a fixative in the preparation of the tissues, and as the couplng agent in the preparation of the immunogen. Following this line we have fixed several brain stem specimens with 0.5% p-benzoquinone; 3% glutaraldehyde; 4% paraformaldehyde plus 0.25% glutaraldehyde and compare the results with tissues fixed in 4% paraformaldehyde. Glutaraldehyde and p-benzoquinone do not produce cyclization of 5-HT but immobilize monoamines in situ. As expected, the antibodies applied according to the PAP technique did not stain the neuronal bodies of the raphe system, known to contain 5-HT when 3–4% glutaraldehyde or 0.5% p-benzoquinone were used. Good staining was obtained with 4% paraformaldehyde alone or with 4% paraformaldehyde plus 0.25% glutaraldehyde.

A quantitative assay of the spot test of Larsson (1981) was devised for measuring in vitro the inhibitory effects of 5-HT, of the 5-HT-BSA complex and of the cyclic derivative, 6-OH-1,2,3,4-tetrahydro-β-carboline. The results confirmed that the avidity of the antiserum is much greater for the cyclic derivatives contained in the 5-HT-BSA complex and for 6-OH-1,2,3,4-tetrahydro-β-carboline than for 5-HT. It is concluded that the formation of a new ring by the lateral chain of 5-HT is responsible of thein-vitro andin the tissue immunoreactivity of the anti-5-HT-antibodies.

Keywords

Formaldehyde Central Nervous System Paraformaldehyde Glutaraldehyde Monoamine 

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Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • S. Peressini
    • 1
  • A. Brusco
    • 1
  • J. Pecci Saavedra
    • 1
  1. 1.Instituto de Biologia Celular, Facultad de MedicinaUniversidad de Buenos AiresBuenos AiresArgentina

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